Can read fragment size be shorter than read length size?
1
0
Entering edit mode
21 months ago
Ivan ▴ 60

I'm trying to calculate the average length of DNA fragment and I'm having some trouble with the concepts of insert size and fragment size.

(The picture for visualization linked here).

I'm using the following command to extract ninth column of a SAM file (the TLEN column)

samtools view -f3 {file} | cut -f 9

The -f3 gives me paired reads mapped in proper pair (per here). My reads are paired, so I get negative values. I take absolute values of those values. The maximum of this array is 500, while the minimum is 89. However, my read length is 101.

How can my minimum value of TLEN column be lesser than my read length?

NGS samtools • 583 views
ADD COMMENT
2
Entering edit mode
21 months ago

How can my minimum value of TLEN column be lesser than my read length?

the read and its' mate overlap.

    R1     ===========> 
    R2            <===========
    TLEN          *****
ADD COMMENT
0
Entering edit mode

Addendum to the question, does the TLEN column then represent the size of DNA fragment or do I have to remove all reads whose TLEN is greater than my read size to get a better estimate of the size of DNA fragment?

ADD REPLY

Login before adding your answer.

Traffic: 2682 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6